Stuck in the Deserts of Tatooine

During the week of November 19th, I was still working on my extractions and began to run gels on them. I figured that if I had my samples incubate overnight in the lysis buffer the fatty membranes and proteins associated with the DNA would have broken down. I also thought I would have seen a DNA pellet form in the -20 degree isopropyl-alcohol after waiting 30 minutes, but I was not able to see anything form. After extracting most of the isopropyl-alcohol from the microcentrifuge-tube, I allowed it to air-dry underneath the hood. I then suspended the DNA in distilled water. My samples underwent electrophoresis and here is the gel:

 This gel held three samples from my three separate DNA extractions. I was not able to see any DNA under the UV light. It was not something I had hoped to see.

Beginning Life on Alderaan

During the week of November 12, I continued trying to perfect my Cacti DNA extraction. This new protocol involved my older protocol to extract DNA from ants. This protocol involved:

• I put 5-15 spines into a microtube and wash with bleach.
• I then decant and rinse the spines twice with distilled water.
• I cut the spines into smaller pieces with nail-cutter scissors.
• I add 90 micro-liters of nuclease free water, 1 micro-liter of 10% SDS, 2 micro-liters of 20% meat tenderizer solution.
• I heated the tube at 75 degrees Celsius for 10 minutes in a water bath and agitated the tube thereafter with a flick of my finger.
• I heated the tube at 95 degree Celsius for 5 minutes in a heating block.
• I agitate the tube with a flick of my finger then allow the tube to incubate in room temperature overnight.
• I take the sample and put it into the centrifuge for 13.2 rpm for 5 min.
• I take the sample out of the centrifuge and remove the supernatant from the top in order to transfer into a new microcentrifuge-tube. 
• I add -20 degrees Celsius isopropyl-alcohol equal to the amount of supernatant and put the tube at -20 degrees Celsius for 30 minutes.
• I centrifuge the sample and collect the supernatant into a new tube.

The heating block used and the other picture shows two samples with -20 degree Celsius isopropyl-alcohol.

Beginning A Trek Into A New Naboo

During the week of November 1st, I began the protocol for extracting DNA from the cholla cacti. I received a piece of cholla from our green house in hopes of extracting DNA and finding clear polymorphic banding after running the DNA sample in a gel. There were various ways on how I began isolating the DNA.

• The first protocol I used consisted of washing 5-15 spines in a microtube and washing them in bleach and decant with a disposable pipet.
• I rinse the spines with distilled water twice and decant.
• I cut the spines with nail-cutter scissors.
• I added 1000 micro-liters of lysis buffer after the spines are cut and incubate for 30 minutes at 65 degrees Celsius. 
• I disrupt the spines by vortex machine and incubate for another 30 minutes.
• I then incubate the sample overnight by allowing it to sit at room temperature.
• I add 5 drops of protease to the microcentrifuge-tube and heat for 10 minutes at 65 degrees Celsius.
• I vortex the sample and put it in the centrifuge at 13.2 rpm for 4.0 minutes.
• I take the sample out of the centrifuge and remove the supernatant from the top in order to transfer into a new microcentrifuge-tube. 
• I add -20 degrees Celsius isopropyl-alcohol equal to the amount of supernatant and put the tube at -20 degrees Celsius for 30 minutes.
• I centrifuge the sample and collect the supernatant into a new tube.